QIAcube HT enables automated mid- to high-throughput nucleic acid purification in 96-well format using silica membrane technology. A variety of QIAcube HT Purification Kits lets users quickly and easily purify DNA, RNA and miRNA from almost any type of sample – including cells, tissues and stool samples, as well as from bacteria and viruses in animal samples. Automated protocols and dedicated accessories and consumables increase reliability and convenience and save valuable time. The system provides the same high-quality results as other trusted QIAGEN purification technologies.

The QIAcube HT is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.

Analysis of genomic DNA purified from human blood.

Human blood from one donor was treated with sodium citrate, heparin, or EDTA. Genomic DNA was purified from a 50 or 200 µl aliquot of the treated blood manually with the QIAamp 96 DNA Blood Kit or on the QIAcube HT using the QIAamp 96 DNA QIAcube HT Kit with the QIAamp 96 DNA QIAcube HT V1.QSP protocol. Purified DNA was eluted in 200 µl. PCR amplification of a 1.2 kb fragment from the single copy Hugl gene was carried out using QIAGEN HotStarTaq DNA Polymerase and 2.5 µl eluate in a total volume of 25 µl. A 1 µl aliquot of each PCR was analyzed on the QIAxcel Advanced with the QIAxcel DNA Screening Kit. M1: pUC18 HaeIII marker; M2: phiX174 HaeIII marker.

Consistent amounts of BVDV particles and Salmonella cells were simultaneously spiked into a variety of animal specimens. Samples were processed in parallel on the QIAcube HT with the cador Pathogen 96 QIAcube HT Kit or manually with the QIAamp cador Pathogen Mini Kit. Co-isolated viral RNA and bacterial DNA were identified using an in-house assay. Mean CT values are shown for [A] BVDV and [B]Salmonella. Small error bars (±1 SD of 3 extraction replicates) indicate high precision for both purification procedures.

Efficient and rapid automated DNA extraction.

The DNeasy mericon 96 QIAcube HT Kit has been adapted for rapid mid- to high-throughput purification of DNA from a variety of raw and processed food matrices on the QIAcube HT instrument. The modified cetyltrimethylammonium bromide (CTAB) extraction procedure allows DNA extraction from food samples in radically shorter time compared with conventional CTAB protocols.

Highly repeatable viral RNA isolation.

Blood samples of various species were spiked with BVDV particles and were frozen at –20°C. Of each sample, 4 replicates were processed on QIAcube HT with the cador Pathogen 96 QIAcube HT Kit in 3 subsequent runs on different days. [A] Viral RNA was identified using an in-house assay, which included [B] an internal amplification control. Error bars represent ±1 SD of 3 replicates. The BVDV CT values for each set of quadruplicates showed a mean relative standard deviation of 0.67 ± 0.43%, and the inter-assay CV for each sample from 3 runs ranged from 0.14 to 0.89% (mean: 0.45%), indicating high repeatability of viral RNA purification on QIAcube HT.

High performance in downstream assays.

Genomic DNA was purified from 200 µl human blood. Samples were processed with the QIAcube HT using the QIAamp 96 DNA QIAcube HT Kit and the QIAamp 96 DNA QIAcube HT V1.QSP protocol, or with the QIAcube using the QIAamp DNA Blood Mini Kit. Quantitative PCR of IL8 was carried out using the QuantiFast Probe Kit with 5 µL eluate in 25 µl. The CT values obtained show even amplification across the whole 96-well plate, indicating no well-to-well variation in DNA recovery with QIAcube HT. CT values were similar between the different purification platforms.

Linear recovery of total RNA from tissue lysates.

Lysates from rat lung tissue stabilized in RNAlater were serially diluted to produce samples containing equivalents of 2.5–25 mg tissue. Total RNA was purified from these samples using the QIAcube HT with the RNeasy QIAcube HT Kit and the RNeasy 96 QIAcube HT total RNA Tissue V1.QSP protocol. Purified RNA was eluted in 100 µl RNase-free water. Optical density measurements show a linear relationship between RNA recovery and amount of starting material.

Purification of DNA from complex food samples.

DNA was purified from orange juice, chocolate, and peanut butter using the QIAcube HT instrument and the DNeasy mericon 96 QIAcube HT Kit or manually using the DNeasy mericon Food Kit. Purified DNA was analyzed by qPCR of the trnL chloroplast tRNA gene using the QuantiFast SYBR® Green PCR Kit. The resulting CT values indicate similar performance from both purification methods. Diluting the input DNA 1:10 for qPCR analysis shows that all purified DNA was also free of PCR inhibitors.

Purification of genomic DNA from tissues.

Genomic DNA was purified from 5 mg rat lung, kidney, spleen, and liver tissue and 200 µl human blood. Samples were processed with the QIAcube HT using the QIAamp 96 DNA QIAcube HT Kit and the QIAamp 96 DNA QIAcube HT V1 QSP protocol or with the QIAcube using the QIAamp DNA Blood Mini Kit (blood) or the QIAamp DNA Mini Kit (tissues). Both platforms showed similar performance in [A] yield and [B] quality of purified DNA for each type of starting material.

Reproducible 96-well RNA purification.

RNA was purified from 5 x 105 Jurkat cells on the QIAcube HT using the RNeasy 96 QIAcube HT Kit with the RNeasy 96 QIAcube HT total RNA cell with DNase V1.QSP protocol. Purified RNA was eluted in 110 µl RNase-free water. Quantitative RT-PCR of the human β-actin mRNA was carried out using the QuantiFast Probe RT-PCR Kit with 5 µl eluate in a total volume of 25 µl. The CT values obtained throughout one 96-well plate show even amplification across the whole plate, indicating no significant well-to-well variation in RNA recovery.

Performance

QIAcube HT provides fast and reliable automated high-throughput nucleic acid purification for use in a variety of sensitive downstream applications. Researchers can easily increase their sample purification throughput and expect the same high-quality results as from other trusted QIAGEN nucleic acid purification solutions.

DNA and RNA from animal samples

QIAcube HT enables reliable purification of nucleic acids from undiluted animal whole blood with minimal risk of silica membrane clogging. Automated processing of samples in 96-well format is precise and repeatable between different purification runs performed on different days, providing not only convenience but also peace of mind (see figure Highly repeatable viral RNA isolation). Co-purification of viral RNA and bacterial DNA is easily automated with QIAcube HT and dedicated cador Pathogen QIAcube HT Kits, and provides the same high-quality nucleic acids as manual processing with the QIAamp cador Pathogen Mini Kit (see figure Co-isolation of viral RNA and bacterial DNA).

DNA from complex food samples

The DNeasy mericon 96 QIAcube HT Kit has been specially adapted for rapid mid- to high-throughput purification of DNA from a variety of raw and processed food matrices on the QIAcube HT instrument. A modified cetyltrimethylammonium bromide (CTAB) extraction procedure allows DNA extraction from all food samples in radically shorter time compared with conventional CTAB protocols (see figure Efficient and rapid DNA extraction from complex food samples). Now users can prepare the same high-quality DNA as from the manual DNeasy mericon Food Kit with a quick and simple automated procedure (see figure Purification of DNA from complex food samples).

Principle

Automated purification on the QIAcube HT is quick and cost-effective and delivers high-quality nucleic acids from 24–96 samples per run. Dedicated QIAcube HT Kits enable purification of nucleic acids from a diverse range of starting materials, including cells, tissues, bacteria, viruses and food material. The QIAcube HT is extremely easy to use, and the intuitive software lets users start a run with just a few mouse clicks.

Five dedicated QIAcube HT purification kits are currently available. The QIAamp 96 DNA QIAcube HT Kit enables automated purification of total DNA from animal blood, cells, and tissue. The QIAamp 96 Virus QIAcube HT Kit provides purification of viral RNA and DNA from a broad range of human sample types including fresh or frozen tissues, blood and other body fluids. The RNeasy 96 QIAcube HT Kit is designed for automated purification of total RNA or total RNA, including miRNA, from animal and human cells and tissue. The cador Pathogen 96 DNA QIAcube HT Kit is for automated purification of viral RNA and DNA and bacterial DNA from animal samples. The DNeasy mericon 96 QIAcube HT Kit is for automated purification of total nucleic acids from a range of raw and processed food sample types. All the plastic consumables required for sample purification runs are conveniently included in the QIAcube HT Plasticware package. Predesigned protocols and instrument run files are easily downloaded from the "Resources" tab on the QIAcube HT catalog page. This maximizes convenience and lets users get up and running quickly in the laboratory.

QIAcube HT is a compact, benchtop instrument with a very small footprint. It fits easily onto most lab benches and can be readily moved into biosafety level laboratories. QIAcube HT comes with a laptop computer for added flexibility.

Advanced instrument safety features, including a translucent hood, help to protect precious samples from environmental contamination. The HEPA filter maintains positive clean air pressure on the worktable underneath the hood and protects precious samples from potential airborne contamination. The UV light provides efficient worktable decontamination and helps to prevent cross-contamination. QIAcube HT ejects used tips through the tip disposal chute into a disposable plastic box located at the side of the instrument. This external tip ejection prevents waste accumulation in the clean workspace. The tip disposal box holder keeps the box positioned correctly under the tip chute. The disposal box also comes with a secure-fitting lid to make waste disposal simple and safe.

Procedure

QIAcube HT provides automated, high-throughput nucleic acid purification for 24–96 samples per run, in increments of 8 samples. The instrument uses a vacuum-driven system to perform the simple and fast bind–wash–elute procedure during sample purification. The vacuum chamber is composed of two separate compartments. This configuration is one of many measures that minimizes sample cross-contamination during purification.

All materials needed for sample purification are placed onto the instrument worktable prior to the run. A 96-well sample block contains the starting sample material that will be lysed and transferred to the filter plate during the run. Easy-to-fill disposable troughs contain all necessary buffers and reagents for sample binding, washing, and elution. Pipet tips are positioned on the right side of the worktable.

Liquid-handling tasks are performed by the 8-channel pipetting head, which provides fast transfer of liquids during the run. The software tracks tip use and internally updates tip availability as the run proceeds. QIAcube HT optimizes the use of plasticware with a smart and economical tip re-use strategy. After washing, the pipetting head moves the 96-well filter plate to the elution compartment, where purified nucleic acids are eluted into the elution microtubes. The included 8-well strip caps securely close the elution microtubes for sample storage.

Software

QIAcube HT Software provides a user-friendly interface for operating the QIAcube HT instrument. The software wizard simulates the instrument worktable and makes it easy to set up and start a run with just a few mouse clicks. The software automatically calculates reagent volumes according to the number of samples, and fully documents every run. The software also allows you to import bar code databases, manage user access, and integrate Laboratory Information Management Systems (LIMS).

These Q Protocols and installation files can only be used with QIAcube HT Operating software version 4.17 and are not compatible with the new QIAcube HT Prep Manager software.

QIAcube HT Prep Manager Software already includes all standard protocols. Additional protocols and kit configurations can be easily installed. Check for available updates under the "Operating Software" section of the "Product Resources" tab.